{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T18:15:23Z","timestamp":1778264123277,"version":"3.51.4"},"reference-count":38,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2024,10,4]],"date-time":"2024-10-04T00:00:00Z","timestamp":1728000000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPq)","award":["142296\/2015-5"],"award-info":[{"award-number":["142296\/2015-5"]}]},{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPq)","award":["408798\/2022-0"],"award-info":[{"award-number":["408798\/2022-0"]}]},{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPq)","award":["231038"],"award-info":[{"award-number":["231038"]}]},{"name":"Fundo Mackenzie de Pesquisa e Inova\u00e7\u00e3o (Mackpesquisa)","award":["142296\/2015-5"],"award-info":[{"award-number":["142296\/2015-5"]}]},{"name":"Fundo Mackenzie de Pesquisa e Inova\u00e7\u00e3o (Mackpesquisa)","award":["408798\/2022-0"],"award-info":[{"award-number":["408798\/2022-0"]}]},{"name":"Fundo Mackenzie de Pesquisa e Inova\u00e7\u00e3o (Mackpesquisa)","award":["231038"],"award-info":[{"award-number":["231038"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: Heterodimer peptides targeting more than one receptor can be advantageous, as tumors can simultaneously express more than one receptor type. For human breast cancer, a promising biological target is tumor angiogenesis through \u03b1v\u03b23 integrin expression. Another promising target is Neuropeptide Y receptors, considering Y1R is overexpressed in 90% of human breast tumors. This article details the development and preclinical evaluation, both in vitro and in vivo, of a novel heterodimer peptide dual-receptor-targeting probe, [99mTc]HYNIC-cRGDfk-NPY, designed for imaging breast tumors. Methods:\u00a0Female BALB\/c healthy mice were used to perform biodistrubution studies and female SCID mice were subcutaneously injected with MCF-7 and MDA-MB-231 tumor cells. [99mTc]HYNIC-cRGDfk-NPY was intravenously administered to the mice, followed by ex vivo biodistribution studies and small-animal SPECT\/CT imaging. Nonspecific tracer uptake in both models was determined by coinjecting an excess of unlabeled HYNIC-cRGDfk-NPY (100 \u00b5g) along with the radiolabeled tracer. Results:\u00a0Imaging and biodistribution data demonstrate good uptake to estrogen receptor-positive (MCF-7) and triple-negative (MDA-MB-231) tumor models. The in vivo tumor uptakes of radiolabeled conjugate were 9.30 \u00b1 3.25% and 4.93 \u00b1 1.01% for MCF-7 and MDA-MB231, respectively. The tumor\/muscle ratios were 5.65 \u00b1 0.94 for the MCF-7 model and 7.78 \u00b1 3.20 for MDA-MB231. Conclusions: [99mTc]HYNIC-cRGDfk-NPY demonstrated rapid blood clearance, renal excretion, and in vivo tumor uptake, highlighting its potential as a tumor imaging agent.<\/jats:p>","DOI":"10.3390\/ph17101328","type":"journal-article","created":{"date-parts":[[2024,10,4]],"date-time":"2024-10-04T12:02:20Z","timestamp":1728043340000},"page":"1328","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Heterobivalent Dual-Target Peptide for Integrin-\u03b1v\u03b23 and Neuropeptide Y Receptors on Breast Tumor"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0846-4785","authenticated-orcid":false,"given":"Aryel H.","family":"Ferreira","sequence":"first","affiliation":[{"name":"MackGraphe-Mackenzie Institute for Research in Graphene and Nanotechnologies, Mackenzie Presbyterian University, S\u00e3o Paulo 01302-907, Brazil"},{"name":"Mackenzie Evangelical College of Paran\u00e1, Mackenzie Presbyterian University, Curitiba 80730-000, Brazil"},{"name":"Nuclear and Energy Research Institute, Instituto de Pesquisas Energ\u00e9ticas e Nucleares da Comiss\u00e3o Nacional de Energia Nuclear\u2014S\u00e3o Paulo (IPEN-CNEN\/SP), S\u00e3o Paulo 05508-000, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6096-3341","authenticated-orcid":false,"given":"Caroline C.","family":"Real","sequence":"additional","affiliation":[{"name":"Department of Nuclear Medicine and PET Center, Aarhus University Hospital, 8200 Aarhus, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Osvaldo","family":"Malafaia","sequence":"additional","affiliation":[{"name":"Mackenzie Evangelical College of Paran\u00e1, Mackenzie Presbyterian University, Curitiba 80730-000, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Li, Z., Aboian, M.S., Zhu, X., and Marquez-Nostra, B. (2022). Clinical Evaluation of Nuclear Imaging Agents in Breast Cancer. Cancers, 14.","DOI":"10.3390\/cancers14092103"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Luining, W.I., Cysouw, M.C.F., Meijer, D., Hendrikse, N.H., Boellaard, R., Vis, A.N., and Oprea-Lager, D.E. (2022). Targeting PSMA Revolutionizes the Role of Nuclear Medicine in Diagnosis and Treatment of Prostate Cancer. Cancers, 14.","DOI":"10.3390\/cancers14051169"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1021\/acs.bioconjchem.0c00617","article-title":"Radiolabeled Peptides and Antibodies in Medicine","volume":"32","author":"Czarnecka","year":"2021","journal-title":"Bioconjug Chem."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Vall-Sagarra, A., Litau, S., Decristoforo, C., W\u00e4ngler, B., Schirrmacher, R., Fricker, G., and W\u00e4ngler, C. (2018). Design, Synthesis, In Vitro, and Initial In Vivo Evaluation of Heterobivalent Peptidic Ligands Targeting Both NPY(Y1)- and GRP-Receptors\u2014An Improvement for Breast Cancer Imaging?. Pharmaceuticals, 11.","DOI":"10.3390\/ph11030065"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Judmann, B., Braun, D., W\u00e4ngler, B., Schirrmacher, R., Fricker, G., and W\u00e4ngler, C. (2020). Current State of Radiolabeled Heterobivalent Peptidic Ligands in Tumor Imaging and Therapy. Pharmaceuticals, 13.","DOI":"10.3390\/ph13080173"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1081","DOI":"10.1007\/s00726-010-0546-y","article-title":"Peptide Heterodimers for Molecular Imaging","volume":"41","author":"Yan","year":"2011","journal-title":"Amino Acids"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cheng, X., H\u00fcbner, R., von Kiedrowski, V., Fricker, G., Schirrmacher, R., W\u00e4ngler, C., and W\u00e4ngler, B. (2021). Design, Synthesis, in Vitro and in Vivo Evaluation of Heterobivalent Sifalin-Modified Peptidic Radioligands Targeting Both Integrin Av \u03923 and the Mc1 Receptor\u2014Suitable for the Specific Visualization of Melanomas?. Pharmaceuticals, 14.","DOI":"10.3390\/ph14060547"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Lindner, S., Rudolf, H., Palumbo, G., Oos, R., Antons, M., H\u00fcbner, R., Bartenstein, P., Schirrmacher, R., W\u00e4ngler, B., and W\u00e4ngler, C. (2021). Are Heterobivalent GRPR- and VPAC1R-Bispecific Radiopeptides Suitable for Efficient in Vivo Tumor Imaging of Prostate Carcinomas?. Bioorg Med. Chem. Lett., 48.","DOI":"10.1016\/j.bmcl.2021.128241"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2726","DOI":"10.1016\/j.apsb.2021.01.004","article-title":"Integrins as Attractive Targets for Cancer Therapeutics","volume":"11","author":"Li","year":"2021","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1002\/jbm.a.36862","article-title":"Recent Progress in Biomedical Applications of RGD-Based Ligand: From Precise Cancer Theranostics to Biomaterial Engineering: A Systematic Review","volume":"108","author":"Alipour","year":"2020","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"228","DOI":"10.2967\/jnumed.116.177048","article-title":"Clinical Translation of a Dual Integrin Av\u03b23- and Gastrin-Releasing Peptide Receptor-Targeting PET Radiotracer, 68Ga-BBN-RGD","volume":"58","author":"Zhang","year":"2017","journal-title":"J. Nucl. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5574","DOI":"10.1021\/acsami.5b00270","article-title":"Neuropeptide Y Y1 Receptors Mediate [Corrected] Targeted Delivery of Anticancer Drug with Encapsulated Nanoparticles to Breast Cancer Cells with High Selectivity and Its Potential for Breast Cancer Therapy","volume":"7","author":"Li","year":"2015","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_13","unstructured":"(2022, June 26). Y1-Mediated Effect of Neuropeptide Y in Cancer|Cancer Research|American Association for Cancer Research. Available online: https:\/\/aacrjournals.org\/cancerres\/article\/61\/11\/4636\/507863\/Y1-Mediated-Effect-of-Neuropeptide-Y-in."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Lin, S.T., Li, Y.Z., Sun, X.Q., Chen, Q.Q., Huang, S.F., Lin, S., and Cai, S.Q. (2021). Update on the Role of Neuropeptide Y and Other Related Factors in Breast Cancer and Osteoporosis. Front. Endocrinol., 12.","DOI":"10.3389\/fendo.2021.705499"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Memminger, M., Keller, M., Lopuch, M., Pop, N., Bernhardt, G., von Angerer, E., and Buschauer, A. (2012). The Neuropeptide Y Y1 Receptor: A Diagnostic Marker? Expression in MCF-7 Breast Cancer Cells Is Down-Regulated by Antiestrogens In Vitro and in Xenografts. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0051032"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.addr.2013.09.007","article-title":"Molecular Imaging for Cancer Diagnosis and Surgery","volume":"66","author":"Hussain","year":"2014","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Freitas, L.F., Ferreira, A.H., Thipe, V.C., Varca, G.H.C., Lima, C.S.A., Batista, J.G.S., Riello, F.N., Nogueira, K., Cruz, C.P.C., and Mendes, G.O.A. (2021). The State of the Art of Theranostic Nanomaterials for Lung, Breast, and Prostate Cancers. Nanomaterials, 11.","DOI":"10.3390\/nano11102579"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1002\/ddr.20265","article-title":"Integrin Alpha(v)Beta(3)-Targeted Cancer Therapy","volume":"69","author":"Liu","year":"2008","journal-title":"Drug Dev. Res."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Florea, A., Mottaghy, F.M., and Bauwens, M. (2021). Molecular Imaging of Angiogenesis in Oncology: Current Preclinical and Clinical Status. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22115544"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Casanovas, O., and Pons-Cursach, R. (2017). Mechanisms of Anti-Angiogenic Therapy. Tumor Angiogenesis, Springer.","DOI":"10.1007\/978-3-319-31215-6_2-2"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1039\/an9952000863","article-title":"Technetium-99m Chelators in Nuclear Medicine. A Review","volume":"120","author":"Hjelstuen","year":"1995","journal-title":"Analyst"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/s40199-017-0179-8","article-title":"99mTc-Radiolabeled GE11-Modified Peptide for Ovarian Tumor Targeting","volume":"25","author":"Rahmanian","year":"2017","journal-title":"DARU J. Pharm. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/S0969-8051(98)00118-8","article-title":"99m-Technetium-Labelled Peptide-HYNIC Conjugates: Effects of Lipophilicity and Stability on Biodistribution","volume":"26","author":"Decristoforo","year":"1999","journal-title":"Nucl. Med. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gaonkar, R.H., Ganguly, S., Dewanjee, S., Sinha, S., Gupta, A., Ganguly, S., Chattopadhyay, D., and Chatterjee Debnath, M. (2017). Garcinol Loaded Vitamin E TPGS Emulsified PLGA Nanoparticles: Preparation, Physicochemical Characterization, in Vitro and in Vivo Studies. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-00696-6"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Brunello, S., Salvarese, N., Carpanese, D., Gobbi, C., Melendez-Alafort, L., and Bolzati, C. (2022). A Review on the Current State and Future Perspectives of [99mTc]Tc-Housed PSMA-i in Prostate Cancer. Molecules, 27.","DOI":"10.3390\/molecules27092617"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Fonseca, I.C.F., Castelo-Branco, M., Cavadas, C., and Abrunhosa, A.J. (2022). PET Imaging of the Neuropeptide Y System: A Systematic Review. Molecules, 27.","DOI":"10.3390\/molecules27123726"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1038\/s41467-022-28510-6","article-title":"Structural Basis of Neuropeptide Y Signaling through Y1 Receptor","volume":"13","author":"Park","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1159\/000486225","article-title":"A Promising Therapeutic Target for Metabolic Diseases: Neuropeptide Y Receptors in Humans","volume":"45","author":"Yi","year":"2018","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.npep.2012.08.005","article-title":"Neuropeptide Y, Peptide YY and Pancreatic Polypeptide in the Gut\u2013Brain Axis","volume":"46","author":"Holzer","year":"2012","journal-title":"Neuropeptides"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/0165-5728(94)90234-8","article-title":"Molecular Cloning of NPY-Y1 Receptor CDNA from Rat Splenic Lymphocytes: Evidence of Low Levels of MRNA Expression and [125I]NPY Binding Sites","volume":"54","author":"Petitto","year":"1994","journal-title":"J. Neuroimmunol."},{"key":"ref_31","first-page":"55","article-title":"Characterization and Molecular Cloning of Vascular Neuropeptide Y Receptor Subtypes in Pig and Dog","volume":"75\u201376","author":"Ekstrand","year":"1998","journal-title":"Regul. Pept."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Chen, W.C., Liu, Y.B., Liu, W.F., Zhou, Y.Y., He, H.F., and Lin, S. (2020). Neuropeptide Y Is an Immunomodulatory Factor: Direct and Indirect. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.580378"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Li, J., Tian, Y., and Wu, A. (2015). Neuropeptide Y Receptors: A Promising Target for Cancer Imaging and Therapy. Regen. Biomater., 2.","DOI":"10.1093\/rb\/rbv013"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1186\/s13550-021-00801-x","article-title":"[99mTc]Tc-Galacto-RGD2 Integrin Av\u03b23-Targeted Imaging as a Surrogate for Molecular Phenotyping in Lung Cancer: Real-World Data","volume":"11","author":"Fu","year":"2021","journal-title":"EJNMMI Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1021\/acsmedchemlett.6b00467","article-title":"Prototypic 18F-Labeled Argininamide-Type Neuropeptide Y Y1R Antagonists as Tracers for PET Imaging of Mammary Carcinoma","volume":"8","author":"Keller","year":"2017","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1016\/j.nucmedbio.2006.09.001","article-title":"[99mTc]HYNIC-RGD for Imaging Integrin Av\u03b23 Expression","volume":"33","author":"Decristoforo","year":"2006","journal-title":"Nucl. Med. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Ahmadpour, S., Noaparast, Z., Abedi, S.M., and Hosseinimehr, S.J. (2018). 99mTc-HYNIC-(Tricine\/EDDA)-FROP Peptide for MCF-7 Breast Tumor Targeting and Imaging. J. Biomed. Sci., 25.","DOI":"10.1186\/s12929-018-0420-x"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s13550-019-0493-x","article-title":"HYNIC and DOMA Conjugated Radiolabeled Bombesin Analogs as Receptor-Targeted Probes for Scintigraphic Detection of Breast Tumor","volume":"9","author":"De","year":"2019","journal-title":"EJNMMI Res."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/17\/10\/1328\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:10:52Z","timestamp":1760112652000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/17\/10\/1328"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,4]]},"references-count":38,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2024,10]]}},"alternative-id":["ph17101328"],"URL":"https:\/\/doi.org\/10.3390\/ph17101328","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,4]]}}}